Multiplex bead suspension array for screening Neisseria gonorrhoeae antibiotic resistance genetic determinants in noncultured clinical samples

J Mol Diagn. 2013 Jan;15(1):116-29. doi: 10.1016/j.jmoldx.2012.08.005. Epub 2012 Nov 14.

Abstract

The increasing threat of antibiotic-resistant Neisseria gonorrhoeae highlights the need for new diagnostic options. A high-throughput multiplex bead suspension array assay was developed for profiling 29 N. gonorrhoeae genomic mutations and 2 plasmid genes conferring resistance to 6 antimicrobial agents: penicillin, ciprofloxacin, cefixime, tetracycline, azithromycin, and spectinomycin. The three steps of this assay include amplification of 12 N. gonorrhoeae chromosomal and plasmid loci, multiplex allele-specific primer extension reaction, and multiplex bead suspension array detection. Antibiotic resistance genetic determinants were identified successfully in 239 cervicovaginal N. gonorrhoeae-positive noncultured swab samples. This molecular assay can be used for detection of gonococci in clinical specimens, molecular typing, mutation profiling, and predictive assessment of N. gonorrhoeae susceptibility to antibiotics without the need for culture.

MeSH terms

  • Alleles
  • Anti-Infective Agents / pharmacology
  • Azithromycin / pharmacology
  • Cefixime / pharmacology
  • Cephalosporins / pharmacology
  • Chromosomes, Bacterial / genetics
  • Ciprofloxacin / pharmacology
  • DNA, Bacterial / genetics
  • Drug Resistance, Bacterial / genetics*
  • Gene Expression Profiling
  • High-Throughput Nucleotide Sequencing / methods*
  • Humans
  • Microbial Sensitivity Tests
  • Mutation
  • Neisseria gonorrhoeae / drug effects*
  • Neisseria gonorrhoeae / genetics*
  • Neisseria gonorrhoeae / growth & development
  • Penicillins / pharmacology
  • Plasmids
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Sequence Analysis, DNA / methods
  • Spectinomycin / pharmacology
  • Tetracycline / pharmacology

Substances

  • Anti-Infective Agents
  • Cephalosporins
  • DNA, Bacterial
  • Penicillins
  • Ciprofloxacin
  • Azithromycin
  • Spectinomycin
  • Cefixime
  • Tetracycline